Stasis technology for storage and transport of natural and engineered tissues

This proposal is focused on developing innovative new cryopreservation techniques employing ice free preservation methods that will lead to the ability to bank complex vascularized tissue and organ grafts Banking of large or complex tissues such as limbs and organs using current tissue banking practices employing conventional cryopreservation by freezing is not possible due to the well known damage caused by intra and extra cellular ice formation The company team has developed vitrification cryopreserved storage in a glassy rather than crystalline ice phase for banking of small tissue samples including blood vessels heart valve tissues and cartilage However scale up to larger tissues more complex tissues and organs needs better ice control during rewarming with less risk of cytotoxicity Our collaborators Professor Duman University of Notre Dame and Professor Ben University of Ottawa have developed natural and synthetic antifreeze compounds respectively that demonstrate benefits in frozen cell systems Duman has developed recombinant insect derived antifreeze proteins and isolation methods for antifreeze glycolipids from several insects and a plant that are very effective at ice control Ben has developed a chemical library that was originally based upon an antifreeze glycoprotein derived from a deep sea teleost We will evaluate the best of these compounds during ice free cryopreservation using a vascular tissue model to determine if they improve ice control during rewarming and reduce the risk of cryoprotectant toxicity by allowing lower cryoprotectant concentrations to be used There are three specific aims in this Phase I feasibility study to identify potentially marketable methods to increase efficacy and minimize tissue damage during cryopreservation of complex natural and engineered tissues In Specific Aim we will prepare of test compounds antifreeze glycolipids from natural sources and synthesize large quantities of synthetic antifreeze compounds A stock of recombinant insect derived antifreeze proteins is already available In Specific Aim we will evaluate the impact of antifreeze compounds individually upon ice free cryopreserved blood vessels and in Specific Aim the best antifreeze supplements will be combined Evaluations will include ice control during ice free vitrification and tissue viability and function assays The technology developed in this proposal will be equally effective for stasis storage of engineered tissue constructs as natural tissues This work will lead to the development of banking by us and other cryobiology groups for internal organs such as livers and hearts vascularized composite allografts such as larynx trachea abdominal wall knee skeletal muscles facial tissues and legs as well as improved methods for critical natural and engineered tissues such as skin heart valves and blood vessels Effective new preservation solutions and protocols are needed as novel complex tissue and organ therapies are developed According to some estimates over of all U S deaths could be prevented or delayed by an organ transplant and broader tissue engineering In the U S organ impairment leads to more deaths than cancer and chronic organ impairment makes up a substantial fraction of these deaths The development of effective cryopreservation methods for large complex tissues and organs would enable upon demand availability of matched banked donor derived tissues and organs Commercialization of antifreeze compound formulations and protocols developed in this proposal will be accomplished by a combination of products cryoprotectant formulations cryopreserved tissue based test systems licensing and clinical processing services to provide important enabling storage technologies for development of multi billion dollar cell and tissue product opportunities